Biological Sample Preparation System Using Pressure Cycling Technology (PCT)

Tao, F.T.; Lawrence, Nathan P.; Miller, Wayne; Li, C.; Tuzmen, P.; Behnke, James; Nakhai, B.; Kakita, Akiyoshi; Christian, Ted J.; Reed, Donald J.; Manak, Mark M.; Schumacher, Richard

doi:10.1007/978-3-662-05613-4_75

Cited by 6 publications

(6 citation statements)

References 1 publication

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Whole mouse brains were disrupted using a pressure cycling technology (PCT) in which the tissues were subjected to repetitive cycles of high and low hydrostatic pressure [6,7]. For this, each of four frozen mouse brains was placed in a PULSE tube with 0.8 mL of 7 M urea, 2 M thiourea, and 1% 3-(4-heptyl)phenyl 3-hydroxypropyl dimethylammonio propane sulfonate (C7BzO).…”

Section: Isolation Of Basic Proteins From Mouse Brainmentioning

confidence: 99%

Tris interference in IEF and 2‐DE

Smejkal¹,

Robinson²

2007

Electrophoresis

View full text Add to dashboard Cite

When dried IPGs are hydrated with protein solutions, the concentration of protein and other ionic constituents is constant throughout the strip. Tris, initially present at a very low concentration, focuses during IEF and accumulates in the gradient at a pH corresponding to its pK(a) at the operative temperature of electrophoresis. Tris focuses more rapidly than many basic proteins, and concentrates into a localized zone of increased conductivity which coincides with a precipitous voltage drop in that vicinity. Basic proteins, already near their pI, are frequently observed to align at the periphery of this zone. Acidic proteins imbibed at the basic end of the gradient must traverse this region before this ionic boundary is formed, or otherwise may fail to migrate to their proper positions in the pH gradient.

show abstract

Section: Isolation Of Basic Proteins From Mouse Brainmentioning

confidence: 99%

Tris interference in IEF and 2‐DE

Smejkal¹,

Robinson²

2007

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…Smejkal et al successfully extracted protein from Escherichia coli using PCT [21]. It has also been used in the extraction of biomolecules such as DNA and RNA from animal and plant tissues, insects, and microbes [20,22,23]. The advantage of pressure cycling is its ability to produce a highly efficient extraction and disruption of cell nuclei from a wide variety of substrates.…”

Section: Introductionmentioning

confidence: 99%

“…Pressure cycling technology for sample preparation (PCT) is a novel method that employs cycles of hydrostatic pressure between ambient and high levels to induce mechanical stress on cells resulting in compromised cellular integrity [19,20]. Application of alternating cycles of hydrostatic pressure between ambient and high levels causes disruption of biomolecular interactions.…”

Section: Introductionmentioning

confidence: 99%

The application of alkaline lysis and pressure cycling technology in the differential extraction of DNA from sperm and epithelial cells recovered from cotton swabs

Nori

McCord

2015

Anal Bioanal Chem

View full text Add to dashboard Cite

This study reports the development of a two-step protocol using pressure cycling technology (PCT) and alkaline lysis for differential extraction of DNA from mixtures of sperm and vaginal epithelial cells recovered from cotton swabs. In controlled experiments, in which equal quantities of sperm and female epithelial cells were added to cotton swabs, 5 min of pressure pulsing in the presence of 0.4 M NaOH resulted in 104 ± 6% recovery of female epithelial DNA present on the swab. Following the pressure treatment, exposing the swabs to a second 5-min alkaline treatment at 95 °C without pressure resulted in the selective recovery of 69 ± 6% of the sperm DNA. The recovery of the vaginal epithelia and sperm DNA was optimized by examining the effect of sodium hydroxide concentration, incubation temperature, and time. Following the alkaline lysis steps, the samples were neutralized with 2 M Tris (pH 7.5) and purified with phenol-chloroform-isoamyl alcohol to permit downstream analysis. The total processing time to remove both fractions from the swab was less than 20 min. Short tandem repeat (STR) analysis of these fractions obtained from PCT treatment and alkaline lysis generated clean profiles of female epithelial DNA and male sperm DNA for 1:1 mixtures of female and male cells and predominant male profiles for mixtures up to 5:1 female to male cells. By reducing the time and increasing the recovery of DNA from cotton swabs, this new method presents a novel and potentially useful procedure for forensic differential extractions.

show abstract

Section: Introductionmentioning

confidence: 99%

“…Pressure cycling technology (PCT) uses a specifically designed device (Barocycler™) and reaction containers (PULSE™ tubes) to apply cycles of hydrostatic pressure to samples [26,27]. PCT provides a simple, fast, effective and reproducible process to release cellular contents from biological samples [26,27,28]. Previously, it has been shown that the use of PCT increased protein yields from E. coli , where PCT extracted 14.2% more total protein than using a standard bead mill [29,30].…”

Section: Introductionmentioning

confidence: 99%

Comparative Proteomic Analysis of Cotton Fiber Development and Protein Extraction Method Comparison in Late Stage Fibers

et al. 2016

View full text Add to dashboard Cite

The distinct stages of cotton fiber development and maturation serve as a single-celled model for studying the molecular mechanisms of plant cell elongation, cell wall development and cellulose biosynthesis. However, this model system of plant cell development is compromised for proteomic studies due to a lack of an efficient protein extraction method during the later stages of fiber development, because of a recalcitrant cell wall and the presence of abundant phenolic compounds. Here, we compared the quality and quantities of proteins extracted from 25 dpa (days post anthesis) fiber with multiple protein extraction methods and present a comprehensive quantitative proteomic study of fiber development from 10 dpa to 25 dpa. Comparative analysis using a label-free quantification method revealed 287 differentially-expressed proteins in the 10 dpa to 25 dpa fiber developmental period. Proteins involved in cell wall metabolism and regulation, cytoskeleton development and carbohydrate metabolism among other functional categories in four fiber developmental stages were identified. Our studies provide protocols for protein extraction from maturing fiber tissues for mass spectrometry analysis and expand knowledge of the proteomic profile of cotton fiber development.

show abstract

Biological Sample Preparation System Using Pressure Cycling Technology (PCT)

Cited by 6 publications

References 1 publication

Tris interference in IEF and 2‐DE

Tris interference in IEF and 2‐DE

The application of alkaline lysis and pressure cycling technology in the differential extraction of DNA from sperm and epithelial cells recovered from cotton swabs

Comparative Proteomic Analysis of Cotton Fiber Development and Protein Extraction Method Comparison in Late Stage Fibers

Contact Info

Product

Resources

About